Cell-free reconstitution of peroxisomal matrix protein import using Xenopus egg extract.
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ABSTRACT:
SUBMITTER: Skowyra ML
PROVIDER: S-EPMC9947420 | biostudies-literature | 2023 Mar
REPOSITORIES: biostudies-literature
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Cell-free reconstitution of peroxisomal matrix protein import using Xenopus egg extract.
Skowyra Michael L ML Rapoport Tom A TA
STAR protocols 20230211 1
Peroxisomes are vital metabolic organelles whose matrix enzymes are imported from the cytosol in a folded state by the soluble receptor PEX5. The import mechanism has been challenging to decipher because of the lack of suitable in vitro systems. Here, we present a protocol for reconstituting matrix protein import using Xenopus egg extract. We describe how extract is prepared, how to replace endogenous PEX5 with recombinant versions, and how to perform and interpret a peroxisomal import reaction ...[more]
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